Process for treating fatigue with calcium glucose-1-phosphate



United States Patent 3,433,869 PROCESS FOR TREATING FATIGUE WITH CALCIUM GLUCOSE-l-PHOSPHATE Roger Brule, Fontenay-sous-Bois, France, assignor to Roussel-UCLAF, Paris, France, a corporation of France No Drawing. Original application July 3, 1964, Ser. No. 380,122. Divided and this application July 13, 1966, Ser. No. 574,519

Claims priority, application France, July 9, 1963, 9 4

U.S. Cl. 424180 Int. Cl. A61k 27/00 3 Claims ABSTRACT OF THE DISCLOSURE A novel salt, calcium glucose-l-phosphate having the formula v ornorr 1|I O\ l O C a .2HO H2) sir/Lao 1 H OH I although the number of water crystallization molecules may vary depending upon the precipitation or crystallization conditions. The invention further relates to novel processes for the preparation of the said calcium salt and to novel compositions for increasing resistance to fatigue. The invention also relates to a method of increasing the resistance of warm-blooded animals to fatigue.

PRIOR APPLICATIONS The present application is a divisional application of my copending, commonly assigned U.S. patent application Ser. No. 380,122, filed July 3, 1964, now abandoned.

PRIOR ART OBJECTS OF THE INVENTION It is an object of the invention to provide the novel compound, calcium glucose-l-phosphate.

It is another object of the invention to provide novel processes for the preparation of calcium glucose-l-phos phate.

It is a further object of the invention to provide novel compositions for increasing resistance to fatigue.

It is an additional object of the invention to provide a novel method of increasing the resistance of Warmblooded animals to fatigue.

These and other objects and advantages of the invention will become obvious from the following detailed description.

THE INVENTION The novel salt of the invention is calcium glucose-1- phosphate which is the calcium salt of the monoester formed by an acid portion of phosphoric acid and the free hydroxy group in the 1-position of glucose in the form of a-d-glucopyranose. The number of molecules of water of crystallization can vary depending upon the method of crystal formation.

The process of the invention for the preparation of calcium glucose-l-phosphate comprises essentially in neutralizing glucose-l-phosphoric acid with calcium hydroxide preferably in an aqueous media and isolating the formed calcium salt by known procedures, such as precipitation by the addition of a water-miscible solvent such as alcohol to the aqueous reaction solution, by lyophilization or by concentration of the aqueous solution to effect crystallization.

A variation of the said process is effecting double decomposition of a salt of glucose-l-phosphoric acid such as an alkali metal salt, ammonium salt or an alkaline earth metal salt other than calcium with a water-soluble calcium salt such as calcium chloride and then recovering calcium glucose-l-phosphate.

Another variant of the process comprises passing an aqueous solution of potassium glucose-l-phosphate through a cation exchange resin to absorb glucose-l-phosphoric acid thereon, eluting glucose-l-phosphoric acid with ammonia to form an aqueous solution of ammonium glucose-l-phosphate and reacting the said salt with a water-soluble calcium salt such as calcium chloride to form calcium glucose-l-phosphate.

The compositions of the invention for increasing resistance to fatigue are comprised of calcium glucose-lphosphate and a major amount of a pharmaceutical carrier. The compositions may be in the form of aqueous solutions for drinkingor injection, in ampules, pills, coated pills, tablets, cachets, aromatic powders, pellets, syrups or suppositories.

The method of the invention for increasing the resistance of Warm-blooded animals to fatigue comprises administering an effective amount of calcium glucose-lphosphate to the animals. The said calcium salt may be administered orally, transcutaneously or rectally. The usual individual oral dose is about 0.50 gm. and the usual daily does is between 1 gm. to 2.5 gm. per day depending upon the method of administration.

In the following examples there are described several preferred embodiments to illustrate the invention. However, it should be understood that the invention is not intended to be limited to the specific embodiments.

Example I.-Preparation of calcium glucose-l-phosphate from the free acid An aqueous solution containing 10 gm. of glucose-1- phosphoric acid was prepared in cc. and While controlling the pH with the aid of a pH-rneter, calcium hydroxide was introduced dropwise into the said solution under agitation until neutralization was complete. This point was attained after the introduction of the stoichiometric amount of calcium hydroxide. Two volumes of ethanol were added to the neutral solution of the resulting calcium salt and the mixture was left standing for a period of 24 hours. The precipitate formed was vacuum filtered, washed in ethanol and vacuum dried to obtain calcium glucose-l-phosphate with a specific rotation [a] =-{78 of water which also was eliminated. The active material fixed on the ion-exchanger was then eluted by percolation at the same flow rate with 1 N ammonium hydroxide.

The rotary power of the efiiuent was determined and all optically active fractions were combined and subjected (c.=4% in water), which occurred in the form of a 5 to a vacuum concentration up to half of their original white or cream-colored, microcrystalline or amorphous volume to expel all the free ammonia. By polarimetric powder. The salt was insoluble in ethanol and most of the means, the exact amount of ammonium glucose-l-phoscommon organic solvents and soluble in water. phate in the solution was determined and the calculated In a pure solution, calcium glucose-l-phosphate exstoichiometric amount of calcium chloride was added to hibited all the conventional reactions of the calcium ion: the solution. After dissolution of the calcium chloride, white precipitate of calcium carbonate upon the addition two volumes of ethanol were added and the solution was of alkali carbonates, white precipitate of calcium sulfate allowed to stand for 24 hours. The precipitate formed was upon the addition of sulfuric acid and soluble sulfates, vacuum filtered, washed with ethanol and vacuum dried white precipitate of calcium oxalate which was insoluble 1 to obtain a product identical to that described in Exin acetic acid and in ammonia by the addition of amample I. monium oxalate. The yield of each of the examples was practically The calcium glucose-I-phosphate solution did not requantitative. spond to the conventional reactions of the mineral phos- PHARMACOLOGICAL STUDY OF CALCIUM phates nor to those of the aldehydic function of the glu- GLUCOSE 1 PHOSPHATE cose, which showed the said product did not undergo hydrolysis. Specifically, the calcium glucose-l-phosphate Resistancfi t0 fatigue-Swimming test solution did not reduce Fehling solution and did not form R weighing between 300 and 360 gm. w Pl d in a el preclplta te of ammomonlagnesla'phosphaee upon a smooth-walled tank filled with water having a temaddltlfm of a fixture of magnesmm e f e The 25 perature of C. The water level was low enough so that ealeulm glucose'l'phoephate dld exhlblt a Posmve the rats could not contact the rim of the tank with their sponse to glucose reactions due to the presence of seve tails. To force the rats to swim constantly, the water was funetfonal groups espeelauy on the Penodlc agitated throughout the test by introducing at the base of oxldatlon macho of Gordon (Anal- Chemthe tank a constant and controlled flow of compressed air. 849)- 30 The rats were considered exhausted when they sank to calclPm glucosejl'phosphme "Y welpletely f f the bottom of the tank and no longer had the strength to lyzable 1n an approxrmately 1 N acid solution upon boillng return to the Surface for 15 minutes with simultaneous formation of glucose Under Such conditions, rats are capable of Swimming and seveeal phosphates' for more than an hour and to reduce the swimming time Analyszs.C H O PCa, 2H O; molecular weight and to hasten the fatigue of the rats, a weight representing 334.2. Calculated: Ca, 11.99%; P, Found: Ca, about 6% of the animals body weight was attached to 1195 15%; 916%- the tail of each. This reduced the swimming time to about Calclum glucose-l-phosphate is not described in the 5 to 6 minutes hterature- 40 For each test, two groups of 10 rats each were used Example II.-Preparation of calcium glucose-l-phosphate and their normal Swimming time 1) was determinedfrom potassium glucose-l-phosphate After 1% hours rest, one group of rats received orally the product being tested and the other group served as the An Selene of 5 of POtaSSIEm glueose'l' control. One hour later or two and a half hours after the phosphate 100 l was Passed by i t through a first swim test, the swim test was repeated and the swim- FP wlth a hmght/fil'imeter ratio elther equal 9 ming time determined was called time after recuperation higher. than and eontemlng of Dowex 1 resm m (T recuperation). Forty-eight hours after the first test, the aeld form percolation was efieeted at a flew of the experiment was repeated except that the control rats volume of eolutlon per 1 of teem Per The from the first test received the test compound and the column was Washed Wlth of l P were 50 treated rats from the first test were the controls. The readded to the pnnelpal effluent The reeultmg eolutlon e cuperation time indicated the recuperative powers of the a pH value between and 2 and treated according rats and any improvement thereof due to the administrato Example I to obtain a product 1dent1cal to that detion of a defatiguing product Senbed m the sale example In Table I, the average T and T times for calcium Example III.Preparation of calcium glucose-l-phosphate -p p j and Vitamin C have been grouped f potassium 1 -1 together. The difference between T and T 1s expressed in percent of increase or decrease and by adding up the An aqueous solution of 10 gm. of potassium glucose-lvariation percentages between the control animals and phosphate in 200 cc. was passed by percolation through a the treated animals, difference in variation in the recupercolumn containing 120 cc. of Dowex 1 resin in the C1- ating periods and, therefore, the defatiguing power of form. The percolation was affected at a flow of 0.5 volume the test compounds was determined. Table I clearly shows of solution per volume of resin per hour and the effluent that calcium glucose-l-phosphate has a definite defatiguing was eliminated. The column was then washed with 200 cc. effect.

TABLE I Percent Product administered Dose, T1 in Till! Percent of ditierence gmJkg. seconds seconds variation in variation Controls 254.33 225 -11.53 Calcium glucose-1-ph0sphate.. 0.250 236.5 236 0.21 11.32 Vitamin C 0. 050 245 253.5 +3.47 15 The toxicity test was effected on mice of the Rockland strain weighing between 18 and 22 gm. and the product was administered in aqueous solution. The results are summarized in Table II.

TABLE II Method of administration Period of observa- DLm in gmJkg.

tion in hours Intravenous 24 1 Intraperitoneal 48 2 Oral 72 2 I claim:

1. A method of increasing the resistance of warmblooded animals to fatigue which comprises administering an effective amount of calcium glucose-l-phosphate to the animals.

2. A method of increasing the resistance of warmblooded animals to fatigue which comprises administering 1 to 2.5 gm. of calcium glucose-l-phosphate to the animals.

3. A method of increasing the resistance of warmblooded animals to fatigue which comprises administering orally 1 to 2.5 gm. of calcium glucose-l-phosphate to the animals.

References Cited UNITED STATES PATENTS 3,114,674 12/1963 Nordmann 167-55 OTHER REFERENCES Chemical Abstracts 19: 2811-2812 (1925 ALBERT T. MEYERS, Primary Examiner.

J. D. GOLDBERG, Assistant Examiner. 

